Lung cancer is the most prevalent cancer worldwide, and radon exposure is ranked as the second risk factor after cigarette smoking. It has been reported that radon induces deoxyribonucleic acid damage and oxidative stress in cells. However, the protein profile and potential biomarkers for early detection of radon-induced lung cancer remain unknown. In this study, we aimed to investigate the effects of intermittent high-dose radon exposure on lung epithelial cells, analyze protein profiles and identify potential biomarkers for diagnosis of radon-related lung cancer. Human lung epithelial cells (A549) were exposed to radon (1000 Bq/m3) for 30 min daily for 7 days. Cell viability was measured using the WST-1 assay, and liquid chromatography-mass spectrometry proteomic analysis was performed. Differentially expressed proteins and gene ontology (GO) enrichment were analyzed. Our findings showed that intermittent high-radon exposure reduced A549 cell viability over time. Proteomic analysis identified proteins associated with stressed-induced apoptosis, mitochondrial adaptation, nuclear integrity and lysosomal degradation. These proteins are related to catabolism, stress response, gene expression and metabolic processes in the biological process of GO analysis. We highlighted specific proteins, including AKR1B1, CDK2, DAPK1, PRDX1 and ALHD2 with potential as biomarkers for radon-related lung cancer. In summary, intermittent high-dose radon exposure affects cellular adaptions of lung epithelial cells including stress-induced apoptosis, mitochondrial dysfunctions and immune regulation. The identified proteins may serve as diagnostic biomarkers or therapeutic targets for radon-related lung cancer.